KR20100015120A - Composition for diagnosing thrombotic thrombocytopenic purpura, thrombotic microangiopathy, or like syndrome thereof, and the method - Google Patents

Abstract

PURPOSE: A composition containing hemoglobin, red blood cell and platelet for diagnosing diagnosis index is provided to early diagnose thrombocytopenic pupura, thrombotic microangiopathy or similar disorder. CONSTITUTION: A composition for diagnosing thrombocytopenic pupura, thrombotic microangiopathy or similar disorder contains hemoglobin, red blood cell and platelet as active ingredient. The similar disorder is second thrombotic microangiopathy or disseminated intravascular coagulation. The diagnosis is performed through calculated index of CA(deviation) value (HDW) of hemoglobin(Hb), Hb in blood, deviation of red blood cell size, red blood cell%(%hyper) having high density, small red blood cell%(%micro) and platelet index.

Description

Composition for diagnosing thrombotic thrombocytopenic purpura, thrombotic microangiopathy, or a similar disease thereof, and a method for preparing the same, and a method for diagnosing thrombotic thrombocytopenic purpura, thrombotic microangiopathy, or like syndrome

The present invention relates to compositions and methods for diagnosing thrombotic thrombocytopenic purpura, thrombotic microangiopathy, or similar diseases thereof.

Generally thrombotic by any one or more of cancer, transplantation, cardiovascular surgery, cardiopulmonary bypass surgery, autoimmune diseases, systemic infections, venom, cyclosporin A, chemotherapy, radiation, ticlopidine, and modified syndromes of hemolytic urethral syndrome Thrombocytopenic purpura, thrombotic microangiopathy, or a similar disease thereof occurs, such a deficiency of ADAMTS 13 (vWF-CP) in the body, causing platelets to attach to very large vWF multimers in the body. Advances blood clots and spreads systemically. As a result, acute kidney injury, anemia, thrombosis, hypoalbuminemia, or hypotension may occur, and the disease may occur at the early stage due to complications of such thrombotic thrombocytopenic purpura, thrombotic microangiopathy, or similar diseases thereof. If it is not confirmed, more than 80% of patients with the disease will die, so prompt diagnosis and treatment of blood infections are required.

To date, the treatment of patients with these disorders has been the treatment of palliatives and antibiotics that slow or relieve symptoms.

However, despite the increasing development of potent antibiotics, infection rates, particularly those that cause thrombotic thrombocytopenic purpura, thrombotic microangiopathy, or similar diseases, are on the rise, but early identification of these diseases is difficult. The difficulty with many powerful therapeutic agents is that they do not adequately describe the timing and application of use due to the lack of accurate and rapid diagnostic methods and means. In addition, the cost of these therapies is quite expensive and would be a huge burden to patients if they were used indiscriminately without the benefit of patients.

The present invention has been devised to solve the above-mentioned necessity and the above problems, and an object of the present invention is to provide a rapid and accurate diagnosis means for thrombotic thrombocytopenic purpura, thrombotic microangiopathy, or similar diseases thereof. .

The present invention includes hemoglobin, erythrocytes and platelets as active ingredients, hemoglobin (Hb), erythrocyte size deviation (RDW), hemoglobin CV (deviation) value (HDW), high density of red blood cells (%) hyper),% small red blood cells (% micro), and an index score for platelets, wherein the index score is 1 point if the hemoglobin level is less than or equal to 12 g / dL, and 0 point greater than 12 g / dL; 1 point if the variation in erythrocyte size is 15% or more, 0 points if less than 15%; 1 point if the CV (deviation) value of hemoglobin is 3.2 g / dL or more, and 0 point less than 3.2 g / dL; 1 point for a high density of red blood cells is 1.9% or more, and 0 points for less than 1.9%; 1 point for small red blood cells is 0.4% or more, 0 points for less than 0.4%; And thrombotic thrombocytopenic purpura, thrombotic microangiopathy, by calculating the number of diagnostic indicators summed up to 1 point if the platelet is 130 × 10 ³ cells / μl or less, and 0 if it is more than 130 × 10 ³ cells / μl. Provided are a disease diagnosis composition for diagnosing a patient having a similar disease thereof and a method for diagnosing the disease using the same.

As can be seen from the above configuration, the diagnostic index score of the present invention is useful for diagnosing patients with thrombotic thrombocytopenic purpura, thrombotic microangiopathy, or similar diseases, and therefore, thrombotic thrombocytopenic necessity for which early diagnosis is essential. It allows for a quick and convenient diagnosis of purpura, thrombotic microangiopathy, or similar diseases thereof.

In order to achieve the above object, the present invention includes hemoglobin, erythrocytes and platelets as active ingredients, hemoglobin (Hb), erythrocyte size deviation (RDW), hemoglobin CV (deviation) value (HDW) of the patient's blood, % Dense red blood cells (% hyper), small sized red blood cells (% micro), and an index score for platelets, where the index score is one point if hemoglobin is less than or equal to 12 g / dL, greater than 12 g / dL Zero point; 1 point if the variation in erythrocyte size is 15% or more, 0 points if less than 15%; 1 point if the CV (deviation) value of hemoglobin is 3.2 g / dL or more, and 0 point less than 3.2 g / dL; 1 point for a high density of red blood cells is 1.9% or more, and 0 points for less than 1.9%; 1 point for small red blood cells is 0.4% or more, 0 points for less than 0.4%; And thrombotic thrombocytopenic purpura, thrombotic microangiopathy, by calculating the number of diagnostic indicators summed up to 1 point if the platelet is 130 × 10 ³ cells / μl or less, and 0 if it is more than 130 × 10 ³ cells / μl. Provided are a disease diagnosis composition for diagnosing a patient having a similar disease thereof.

In the composition of the present invention, when the diagnostic indicator score is 6, the death rate due to thrombotic thrombocytopenic purpura, thrombotic microangiopathy, or a similar disease thereof is increased. Thrombocytopenic purpura, thrombotic microangiopathy, or similar diseases thereof. Examples of the similar disease include, but are not limited to, secondary thrombotic microangiopathy or disseminated intravascular coagulation.

In addition, the present invention includes hemoglobin, erythrocytes and platelets as active ingredients, hemoglobin (Hb), erythrocyte size deviation (RDW), hemoglobin CV (deviation) value of hemoglobin (HDW), high density of red blood cells ( % hyper),% small red blood cells (% micro), and an index score for platelets, wherein the index score is 1 point if the hemoglobin level is 12 g / dL or less, and 0 point greater than 12 g / dL; 1 point if the variation in erythrocyte size is 15% or more, 0 points if less than 15%; 1 point if the CV (deviation) value of hemoglobin is 3.2 g / dL or more, and 0 point less than 3.2 g / dL; 1 point for a high density of red blood cells is 1.9% or more, and 0 points for less than 1.9%; 1 point for small red blood cells is 0.4% or more, 0 points for less than 0.4%; And thrombotic thrombocytopenic purpura, thrombotic microangiopathy, by calculating the number of diagnostic indicators summed up to 1 point if the platelet is 130 × 10 ³ cells / μl or less, and 0 if it is more than 130 × 10 ³ cells / μl. A method of diagnosing a patient with their similar disease is provided.

In the method of the present invention, when the diagnostic indicator score is 6, the death rate due to thrombotic thrombocytopenic purpura, thrombotic microangiopathy, or a similar disease thereof is increased. Thrombocytopenic purpura, thrombotic microangiopathy, or similar diseases thereof.

Hereinafter, the present invention will be described.

The inventors of the present invention made a diagnostic index score, and when the score is 6, it was confirmed that it is associated with thrombotic thrombocytopenic purpura, thrombotic microangiopathy, or similar diseases thereof. The present invention analyzed blood infection at Konyang University Hospital in Daejeon, Korea from 2005 to 2007 using the diagnostic index score value.

In the present invention, the diagnostic index score means the sum of the index scores for hemoglobin (Hb), RDW, HDW,% hyper,% micro and platelets in blood. The index scores of the above figures are 1 point for hemoglobin levels in the blood composition of the patient below 12 g / μs, 0 points over 12 g / μs, 1 point for RDW above 15%, 0 points below 15%, and HDW. Is 1 point if 3.2 g / ㎗ or more, 0 point if less than 3.2 g / ㎗, 1 point if% hyper is more than 1.9%, 0 point if less than 1.9%, 1 point if less than 0.4%, less than 0.4% It is defined as 0 point and 1 point if the platelet is 130 × 10 ³ cells / μl or less, and 0 point if it is more than 130 × 10 ³ cells / μl.

 The data show values for blood outside the normal range and diagnosed with thrombotic thrombocytopenic purpura, thrombotic microangiopathy, or similar diseases, as in FIG. As a result, the present invention enables rapid and convenient diagnosis of thrombotic thrombocytopenic purpura, thrombotic microangiopathy, or similar diseases in which early mortality is very high when early treatment fails.

The inventors of the present invention made diagnostic indicator scores using an automated blood analyzer ADVIA 120 (Bayer Diagnostics Tarrytown, New York, USA). From 2005 to 2007, a general blood test was performed on a number of patients using ADVIA 120.

As a result of analyzing the blood test results of the plurality of patients, the patient showing the data as shown in Figure 1, that is the blood composition of hemoglobin level (Hb) 12 g / ㎗ or less, RDW 15% or more, HDW 3.2 g / ㎗ or more,% hyper 1.9 Twelve patients with a diagnostic index score of 6% or more,% micro 0.4% or more, and platelets of 130 × 10 ³ cells / μL or less all had thrombotic thrombocytopenic purpura, thrombotic microangiopathy, or the like. The disease was identified as having a disease, and patients with a diagnostic index score of 6 were found to have a disease other than thrombotic thrombocytopenic purpura, thrombotic microangiopathy, or a similar disease thereof.

In conclusion, the diagnosis score was 6 and the severity of the symptom was severe and the mortality rate was high.However, even when the diagnosis score was 6, the white blood cell count was normal, so there was no correlation between the diagnostic score and the white blood cell count. Did. Therefore, without early treatment, it is thought that the diagnostic index score may be used as a useful indicator for early diagnosis of thrombotic thrombocytopenic purpura, thrombotic microangiopathy, or similar diseases with a high mortality rate.

Hereinafter, the present invention will be described in more detail with reference to non-limiting examples.

Example  One

Hemoglobin method ( Hemoglobin Method )

Samples and ADIVIA 120 HGB reagent are placed in a hemoglobin reaction chamber and mixed, erythrocytes are lysed and hemoglobin is released from the cells. Afterwards, the iron component of the heme group in hemoglobin is oxidized to change from ferrous state to ferric state. The ferric state turns iron into cyanide from the ADIVIA 120 HGB reagent to produce the reaction product. It was optically colorimetrically measured at 546 nm to measure the degree of light transmission of hemoglobin. Thus, drawings of HGB Trans and the like were obtained. The above procedure was carried out using an automatic blood analyzer ADVIA 120 (Bayer Diagnostics Tarrytown, New York, USA) and in accordance with the instructions described on the device.

Erythrocyte / platelet method ( RBC Of Platelet Method )

Erythrocytes and platelets are spheronized to a uniform volume by the sodium dodecyl sulfate (SDS) and glutaraldehyde components of ADIVIA 120 RBC / PLT reagent, and then the erythrocytes and platelets are fixed. The erythrocytes were passed through a flowcell to measure the volume and hemoglobin concentration of erythrocytes through low-angle light scatter and high-angle light scatter of each erythrocyte. In addition, the platelets are passed through the flowcell to amplify the low-angle light scatter signal of each platelet 30 times and to amplify the high-angle light scatter signal 12 times to increase the volume and refractive index of the platelets. ) Was measured. Thus, drawings of RBC HC, RBC Volume, RBC V / HC, and Platelet Vol were obtained. The above procedure was carried out using an automatic blood analyzer ADVIA 120 (Bayer Diagnostics Tarrytown, New York, USA) and in accordance with the instructions described on the device.

Reticulocyte method ( Reticulocyte Method )

Erythrocytes and platelets were spheroidized in a uniform volume by the zwitterionic detergent component of the ADIVIA 120 autoRETIC reagent, and then the reticulocytes were oxazine 750 (Oxazine 750, cationic dye) of the ADIVIA 120 autoRETIC reagent. Stain differently depending on the RNA content. A certain amount of sample from the reticulocyte reaction chamber was passed through a flowcell to obtain cell size, concentration of hemoglobin, and RNA, respectively, via low-angle light scatter, high-angle light scatter, and absorbance. The content was measured. Stained reticulocytes absorb more light than mature red blood cells. Thus, drawings of Retic CH and Retic Scatter Abs were obtained. The above procedure was carried out using an automatic blood analyzer ADVIA 120 (Bayer Diagnostics Tarrytown, New York, USA) and in accordance with the instructions described on the device.

Peroxidase  Way( Peroxidase Method )

Dissolve red blood cells using perox reagent, stain and fix white blood cells, and then pass the stained cells through a flow cell using a Tungsten lamp as a light source, measuring the white blood cell size through scattered light of each white blood cell, and measuring the absorbance according to the peroxidase staining intensity. Each white blood cell was differentiated by. This obtained the drawing of Perox et al. The above procedure was carried out using an automatic blood analyzer ADVIA 120 (Bayer Diagnostics Tarrytown, New York, USA) and in accordance with the instructions described on the device.

Bassopil Of Lobe diagram  Way( Basophil Of lobularity Method )

By completely dissolving erythrocytes and platelets using the inherent pH value of blood cells, all kinds of leukocytes except basophil are removed from the cell membrane and cytoplasm, leaving only the nuclei. Cell size and mesenchyme were measured and divided into monocytes and polymorphonuclear cells. This obtained the drawing of Baso et al. The above procedure was carried out using an automatic blood analyzer ADVIA 120 (Bayer Diagnostics Tarrytown, New York, USA) and in accordance with the instructions described on the device.

1 is a diagram showing the RBC V / HC for the blood, the case in the center of the rectangle corresponds to the normal range. The horizontal axis represents the density of red blood cells and the vertical axis represents the size of red blood cells. The disease of the present invention may be diagnosed as a patient with thrombotic thrombocytopenic purpura, thrombotic microangiopathy, or a similar disease in the lower right part of this figure, that is, a patient with high density and small red blood cells. have.

The data of the patients shown in FIGS. 2A and 2B show that although the red blood cells are small and high in density, since the white blood cell count is normal and the WBC percentage is normal, everyone can misunderstand these patients as normal. However, the diagnostic index score for these patients is six. Eventually these patients died of thrombotic thrombocytopenic purpura, thrombotic microangiopathy, or similar diseases. Thus, in particular, the present invention shows that it is useful for early diagnosis of patients with normal leukocyte count and suffering from thrombotic thrombocytopenic purpura, thrombotic microangiopathy, or similar diseases thereof. In Figures 2A and 2B, the definitions of terms for each data are as follows:

Header-WB: patient information;

Routine Retic: general measurement data of reticulocytes of ADVIA 120;

Routine CBC: Complete Blood Count (CBC) measurement of ADVIA 120 as Hemoglobin (Hb), Hematocrit (Hct), Mean Corpuscular Volume (MCV), Mean Hemoglobin Concentration (Mean Corpuscular Hemoglobin Concentration, MCHC), mean hemoglobin amount (Mean Corpuscular Hemoglobin (MCH)), red cell distribution width (RDW), hemoglobin distribution width (HDW) and platelet count (Platelet count);

Routine WBC Differential: data for neutrophils, lymphocytes, monocytes, eosinophils, basophils and large unstained cells (LUC) as leukocyte differential calculations;

Additional Routine Parameters: additional variables;

Morphology Flags: data in which +, ++, or +++ is indicated when abnormalities in the shape of white blood cells or red blood cells are suspected;

Sample / System Flags: data in +, ++, or +++ if the samples are wrong or if there is any suspicion of equipment failure;

Perox: measurement data for peroxidase staining and cell size of each cell by ADVIA 120;

Baso: measurement data for nucleus morphology and cell size by ADVIA 120;

Retic Scatter Abs: measurement data on cell maturity and cell size by ADVIA 120;

RBC HC: distribution of RBCs according to hemoglobin concentration of RBCs;

RBC Volume: distribution of RBC over cell volume;

Retic CH: distribution of mature RBCs and reticulocytes according to Hb content (cellular Hb);

Platelet Vol: distribution by platelet volume;

HGB Trans: Optical colorimetric data at 546 nm light of hemoglobin.

Claims (6)

It contains hemoglobin, erythrocytes and platelets as active ingredients, hemoglobin (Hb), erythrocyte size deviation (RDW), hemoglobin CV (HDW), high density red blood cells (% hyper), Small red blood cell% (% micro), and an index score for platelets, wherein the index score is 1 point if the hemoglobin level is less than or equal to 12 g / dL, and 0 point greater than 12 g / dL; 1 point if the variation in erythrocyte size is 15% or more, 0 points if less than 15%; 1 point if the CV (deviation) value of hemoglobin is 3.2 g / dL or more, and 0 point less than 3.2 g / dL; 1 point for a high density of red blood cells is 1.9% or more, and 0 points for less than 1.9%; 1 point for small red blood cells is 0.4% or more, 0 points for less than 0.4%; And thrombotic thrombocytopenic purpura, thrombotic microangiopathy, by calculating the number of diagnostic indicators summed up to 1 point if the platelet is 130 × 10 ³ cells / μl or less, and 0 if it is more than 130 × 10 ³ cells / μl. A disease diagnostic composition for diagnosing a patient having a similar disease thereof. The method of claim 1, The diagnostic indicator score is 6, the composition for diagnosing diseases for diagnosing patients with thrombotic thrombocytopenic purpura, thrombotic microangiopathy, or similar diseases thereof. The method according to claim 1 or 2, The similar disease is secondary thrombotic microangiopathy or disseminated intravascular coagulation. Composition. It contains hemoglobin, erythrocytes and platelets as active ingredients, hemoglobin (Hb), erythrocyte size deviation (RDW), hemoglobin CV (HDW), high density red blood cells (% hyper), Small red blood cell% (% micro), and an index score for platelets, wherein the index score is 1 point if the hemoglobin level is less than or equal to 12 g / dL, and 0 point greater than 12 g / dL; 1 point if the variation in erythrocyte size is 15% or more, 0 points if less than 15%; 1 point if the CV (deviation) value of hemoglobin is 3.2 g / dL or more, and 0 point less than 3.2 g / dL; 1 point for a high density of red blood cells is 1.9% or more, and 0 points for less than 1.9%; 1 point for small red blood cells is 0.4% or more, 0 points for less than 0.4%; And thrombotic thrombocytopenic purpura, thrombotic microangiopathy, by calculating the number of diagnostic indicators summed up to 1 point if the platelet is 130 × 10 ³ cells / μl or less, and 0 if it is more than 130 × 10 ³ cells / μl. Method of diagnosing patients with their similar disease. The method of claim 4, wherein The diagnostic indicator score is 6, a method for diagnosing a patient with thrombotic thrombocytopenic purpura, thrombotic microangiopathy, or a similar disease thereof. The method according to claim 4 or 5, The similar disease is secondary thrombotic microangiopathy or disseminated intravascular coagulation. Way.

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